Detection of Anaplasma phagocytophilum in Brazilian dogs by real-time polymerase chain reaction

Abstract

Anaplasma phagocytophilum was detected in dogs from Brazil in the municipalities of Seropédica and Itaguaí, Rio de Janeiro state, by real-time polymerase chain reaction (PCR) using SYBR Green to detect the amplification. Of 253 samples, 18 (7.11%) were positive, with a threshold cycle (Ct) ranging from 31 to 35 cycles. The PCR product from a positive sample was cloned and sequenced. The sequence obtained demonstrated 100% identity with other A. phagocytophilum sequences published in the GenBank database. The analytical sensitivity of RT-PCR using SYBR Green system was able to detect 3 plasmid copies when defined numbers of plasmid copies containing 122 base pairs from the msp2 gene were used. The assay was considered specific when DNA from bacteria (Anaplasma platys, Anaplasma marginale, Ehrlichia canis, Neorickettsia risticii, Rickettsia rickettsii) closely related to A. phagocytophilum was placed in the reaction. These results demonstrate that the canine granulocytic anaplasmosis agent is present in regions in which dogs could be a source of infection for tick vectors. The current study reports the detection of A. phagocytophilum, a zoonotic agent responsible for Human granulocytic anaplasmosis, in Brazilian dogs.

Detecção de Theileria equi e Babesia caballi e anticorpos anti-Ehrlichia spp. em equídeos do Pantanal Mato-Grossense, Brasil

“…nas amostras testadas pela PCR. Ao contrário de N. risticii, espécies filogeneticamente próximas ou idênticas a A. phagocytophilum já foram detectadas em cães, cervídeos e aves no Brasil (Santos et al, 2011;Machado et al, 2012;Secchi et al, 2012 (Salvagni et al, 2010). No presente estudo, 39,6% dos animais foram reagentes a antígenos de Ehrlichia spp.…”

Molecular and serological detection of tick-borne pathogens in dogs from an area endemic for Leishmania infantumin Mato Grosso do Sul, Brazil

“…Although dogs infected with A. platys and A. phagocytophilum usually present mild symptoms or are asymptomatic (HARRUS et al, 1997;KOHN et al, 2008;RAR;GOLOVLJOVA, 2011), some cases may be serious or fatal, particularly when co-infections are involved (DANTAS-TORRES, 2008). The confirmation of the occurrence of E. ewingii ) and A. phagocytophilum (SANTOS et al, 2011) in dogs from Minas Gerais and Rio de Janeiro, respectively, showed that other Anaplasmataceae agents circulate among dogs in Brazil, although the vector tick species involved in these transmission routes remains unknown.…”

“…Anaplasma platys, a bacterium that parasitizes platelets, causing canine cyclic thrombocytopenia (COHN, 2003), has also been reported in dogs in Brazil (DAGNONE et al, 2009). Furthermore, A. phagocytophilum also has been detected molecularly in dogs in the state of Rio de Janeiro, Brazil (SANTOS et al, 2011). Although dogs infected with A. platys and A. phagocytophilum usually present mild symptoms or are asymptomatic (HARRUS et al, 1997;KOHN et al, 2008;RAR;GOLOVLJOVA, 2011), some cases may be serious or fatal, particularly when co-infections are involved (DANTAS-TORRES, 2008).…”

Migratory and Carnivorous Birds in Brazil: Reservoirs forAnaplasmaandEhrlichiaSpecies?

“…Until now, E. chaffeensis and A. phagocytophilum have only been detected in marsh deer (Blastocerus dichomotus) (Machado et al 2006), and dogs (Santos et al 2011) in Brazil, respectively. E. chaffeensis DNA found in the hawk in our study was closely related to E. chaffeensis found in Brazilian marsh deer (Machado et al 2006) in the 16S rRNA phylogenetic tree.…”

“…In Brazil, Ehrlichia chaffeensis and Anaplasma phagocytophilum have been detected in Brazilian marsh deer (Blastocerus dichotomus) (Machado et al 2006) and dogs (Santos et al 2011), respectively. Regarding the occurrence of ehrlichiosis among humans in Brazil, only serological studies have been reported (Calic et al 2004;Costa et al 2005).…”